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Progress in Natural Science:Materials International 2005年第11期15卷 988-992页

作者： TANG Fang, LIANG Pei and GAO Xiwu (Department of Entomology, china agricultural university, beijing 100094. china) Department of Entomology China Agricultural University Beijing 100094 China

The tissue-specific expression of glutathione S-transferases (GSTs) in the cotton bollworm and the expression level induced by 2-tridecanone and quercetin were examined using the methods of biochemistry and the quantitative PCR. The relative expression level of GST mRNA was unanimous with the GSTs activity conjugaging with 1-chloro-2, 4-dimitro-benzene (CDNB) in fat bodies, midguts, heads and integuments of cotton bollworms. The GSTs activity in fat bodies was the highest, then midguts, heads and integuments in turn, which was in consistent with the relative expression level of GST mRNA. The specific activity of GSTs and the relative expression level of GST mRNA could be significantly induced by 2-tridecanone and quercetin, and after the induction the order of the GSTs activity and the relative expression level of GST mRNA in the above four tissues in cotton bollworms was not different from the control. The induction of GSTs by 2-tridecanone was stronger than by quercetin in all four tissues, which was in accordance with the relative expression level of GST mRNA. It suggested that the increase of GSTs activity induced by plant allelochemicals was associated with the elevated expression of GST mRNA in cotton bollworms.

关键词： Helicoverpa armigera glutathione S-transferase tissue-specific expression plant allelochemicals.

Aberrant histone H4 acetylation in dead somatic cell-cloned calves

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Progress in Natural Science:Materials International 2008年第7期18卷 813-818页

作者： Lei Zhang a,b,*, Shaohua Wang a, Qiang Li a, Xiangdong Ding c, Yunping Dai a, Ning Li a,* a State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100094. china b beijing Gene & Protein Biotechnology Co. LTD, beijing 100094. china c College of Animal Science and Technology, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology China Agricultural University Beijing 100094 China beijing Gene & Protein Biotechnology Co. LTD Beijing 100094 China College of Animal Science and Technology China Agricultural University Beijing 100094 China

In somatic cell-cloned animals, inefficient epigenetic reprogramming can result in an inappropriate gene expression and histone H4 acetylation is one of the key epigenetic modifications regulating gene expression. In this study, we investigated the levels of histone H4 acetylation of 11 development-related genes and expression levels of 19 genes in lungs of three normal control calves and nine aber- rant somatic cell-cloned calves. The results showed that nine studied genes had decreased acetylation levels in aberrant clones (p 0.05). Whereas 13 genes had significantly decreased expression (p 0.05), and only one gene had higher expression level in clones (p < 0.05). Furthermore, FGFR, GHR, HGFR and IGF1 genes showed lowered levels of both histone H4 acetylation and expression in aberrant clones than in controls, and the level of histone H4 acetylation was even more lowered in aberrant clones than those in controls. It was suggested that the lower levels of histone H4 acetylation in aberrant clones caused by the previous memory of cell differentiation might not support enough chromatin reprogramming, thus affecting appropriate gene expressions, and growth and development of the cloned calves. To our knowledge, this is the first study on how histone H4 acetylation affects gene expression in organs of somatic cell-cloned calves.

关键词： Somatic cell-cloned calves Histone H4 acetylation Gene expression Lungs

Genetic analysis on 3'- terminal flanking region of uncoupling protein 3 in different pig breeds

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Chinese Science Bulletin 2002年第18期47卷 1541-1543页

作者： FANG Meiying, ZHAO Xingbo, LI Ning & WU ChangxinCollege of Animal Science and Technology, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology, china agricultural university, beijing 100094. china College of Animal Science and Technology China Agricultural University Beijing China State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China

The 3’-terminal flanking region of porcine uncoupling protein 3 (UCP3) was cloned, the sequence data revealed 15 nucleotide substitutions among Landrace and three Chinese native pig breeds named Neijiang, Minpig and Erhualian. The continuous 9 polymorphic sites were checked by PCR-RFLP, the results indicated that Erhualian had extraordinary gene frequency, presented most significant difference by x2 test compared with Landrace, Largewhite, Neijiang and Minpig respectively, significant level compared with Meishan; and Meishan also had significant difference compared with Landrace and Minpig respectively. These results can be concluded that Taihu pigs have special genetic characteristics among pig breeds.

关键词： pig uncoupling protein gene clone PCR-RFLP.

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Tn5-mutagenesis and identification of atr operon and trpE gene responsible for indole-3-acetic acid synthesis in Azospirillum brasilense Yu6.

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Progress in Natural Science:Materials International 2006年第4期16卷 366-372页

作者： XIE Baoen and CHEN Sanfeng State Key Laboratory for Agrobiotechnology, College of Biological Sciences, china agricultural university, beijing 100094. china State Key Laboratory for Agrobiotechnology College of Biological Sciences China Agricultural University Beijing 100094 China

To bring more information about synthesis of indole-3-acetic acid (IAA) from Azospirillum brasilense, a Tn5-insertion library of A. brasilense Yu6. was constructed and subjected to screening for IAA producing mutants. Two mutants with decreased IAA levels, named as A3 and A24, were isolated. The sequence analysis of loci tagged showed that the Tn5-106.a was located in the atrA gene encoding GntR family transcriptional regulator and trpE gene encoding component I of anthranilate synthase respectively. At the same time. atrB encoding phosphotransferase and atrC encoding aminotransferase were cloned downstream the atrA gene and atrA, atrB and atrC were clustered in an operon. Mutagenesis and complementation studies showed that atrA and atrC were involved in IAA synthesis. IAA levels of trpE mutant and wild-type strain could be improved by adding anthranilate into the medium.

关键词： indole-3-acetic acid (IAA) IAA mutants Azospirillum brasilense atrABC cluster trpE Tn5 transposon.

Carbon isotope ratios of C_(4) plants in loess areas of North china

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Science china Earth Sciences 2006年第1期49卷 97-102页

作者： WANG Guoan1,2,3, HAN Jiamao 2, ZHOU Liping3, XIONG Xiaogang4, TAN Ming2, WU Zhenhai5 & PENG Jun6.1. Department of Plant Nutrition, china agricultural university, beijing 100094. china 2. Institute of Geology and Geophysics, Chinese Academy of Sciences, beijing 100029, china 3. College of Environmental Sciences, MOE Laboratory for Earth Surface Processes, Peking university, beijing 100871, china 4. Institute of Botany, Chinese Academy of Sciences, beijing 100093, china 5. Northwest Sci-Tech university of Agriculture and Forestry, Yangling 712100, china 6. Network center, china agricultural university, beijing 100094. china 1. Department of Plant Nutrition China Agricultural University Beijing 100094 China 2. Institute of Geology and Geophysics Chinese Academy of Sciences Beijing 100029 China 3. College of Environmental Sciences MOE Laboratory for Earth Surface Processes Peking University Beijing 100871 China 4. Institute of Botany Chinese Academy of Sciences Beijing 100093 China 5. Northwest Sci-Tech university of Agriculture and Forestry Yangling 712100 China 6. Network center China Agricultural University Beijing 100094 China

Carbon isotope ratios (δ^(13)C) of 89 C_4 plant samples were determined fromthe loess area in North china. δ^(13)C values vary between -10.5per thousand and -14.6.er thousandwith a mean of -12.6.er thousand. Along a precipitation gradient from the semi-moist area to thesemiarid area, then to the arid area, the δ^(13)C values of C_4 plants show a slight decreasingtrend. The δ^(13)C values of C_4 plants in the dry season are found lower than those in the wetseason. These trends are opposite to those observed for C_3 species.

关键词： C4 plants, carbon isotope ratios, loess area.

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Types of voltage-dependent calcium channels involved in high potassium depolarization-induced amylase secretion in the exocrine pancreatic tumour cell line AR4-2J

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Cell Research 1998年第1期8卷 23-31页

作者： CUI ZONG JIE (beijing agricultural university Faculty of Biological Sciences beijing 100094. china) beijing agricultural university Faculty of Biological Sciences Beijing 100094 China

In the perifused fura-2 loaded exocrine pancreatic acinar cell line AR4-2J pulses of high potassium induced repetitive increases in intracellular calcium. Attached cells when stimulated with high potassium secreted large amount of amylase. High potassium-induced secretion was dependent both on the concentration of potassium and duration of stimulation. High potassium induced increases in intracellular calcium were inhibited by voltage-dependent calcium channel antagonists with an order of potency as follows: nifedipine > ω-agatoxin IVA > ω-conotoxin GVIA. In contrast, the L-type calcium channel antagonist nifedipine almost completely inhibited potassium-induced amylase secretion, whereas the N-type channel antagonist ω-conotoxin GVIA was without effect. The P-type channel antagonist ω-agatoxin IVA had a small inhibitory effect, but this inhibition was not significant at the level of amylase secretion. In conclusion, the AR4-2J cell line possesses different voltage-dependent calcium channels (L, P,N) with the L-type predominantly involved in depolarization induced amylase secretion.

关键词： AR4-2J pancreatic acinar cells amylase secretion calcium channels

Effect of Soil Moisture on Release of Low-Molecular Weight Organic Acids in Root Exudates and the Accu-mulation of Iron in Root Apoplasm of Peanut

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Pedosphere 2000年第4期10卷 333-338页

作者： ZHENG YI and ZHANG FUSUO (Department of Plant Nutrition, china agricultural university, beijing,100094.china) 中国农业大学北京 100094

A three-compartments rhizobox was designed and used to study the low-molecular-weight organic acids in root exudates and the root apoplastic iron of "lime-induced chlorosis" peanut grown on a calcareous soil in relation to different soil moisture conditions. Results showed that chlorosis of peanuts developed under condition of high soil moisture level (250 g kg-1), while peanuts grew well and chlorosis did not develop when soil moisture was managed to a normal level (150 g kg-1). The malic acid, maleic acid and succinic acid contents of chlorotic peanut increased by 108.723, 0.029 and 22.446.ig cm-2, respectively, compared with healthy peanuts. The content of citric acid and fumaric acid also increased in root exudates of chlorotic peanuts. On Days 28 and 42 of peanut growth, the accumulation of root apoplastic iron in chlorotic peanuts was higher than that of healthy peanuts. From Day 28 to Day 42, the mobilization percentages of chlorotic peanuts and healthy peanuts to root apoplastic iron were almost the same, being 52.4% and 52.8%, respectively, indicating that the chlorosis might be caused by the inactivation of iron within peanut plant grown on a calcareous soil under high soil moisture conditions.

关键词： apoplastic iron peanut root exudates soil moisture

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Migration and population genetics of the grain aphid Macrosiphum miscanti (Takahashi) in relation to the geographic distance and gene flow

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Progress in Natural Science:Materials International 2005年第11期15卷 1000-1004页

作者： GUO Wei, SHEN Zuorui, LI Zhihong and GAO Lingwang (College of Agronomy and Biotechnology, china agricultural university, beijing 100094. china) College of Agronomy and Biotechnology China Agricultural University Beijing 100094 China

The population genetics of the grain aphid Macrosiphum miscanti (Takahashi) is analyzed by microsatellite markers. Samples collected from 15 locations in china have been examined at 5 polymorphic microsatellite loci. Overall, genetic diversity displays a relation between the migration and gene flow in the grain aphid; a free and frequent gene flow is found in the eastern populations, and gene isolation occurs in the two western populations, especially Datong population and Guiyang population. The natural barriers may present an insurmountable obstacle preventing gene flow and aphid migration. However, a spatial genetic differentiation between populations is correlated with their geographical separation, indicating the geographic differentiation may play an important role in shaping the genetic structure of M. miscanti populations. In addition, most populations of grain aphids are out of Hardy-Weinberg equilibrium and there is het-erozygote deficit. Based on F statistics, the average genetic differentiation among different geographical populations is relatively low. It seems that the long distance migration of the grain aphid may enhance gene flow and decrease genetic differentiation among different populations .

关键词： Macrosiphum miscanti microsatellite gene flow migration.

A five-fold pig bacterial artificial chromosome library:a resource for positional cloning and physical mapping

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Progress in Natural Science:Materials International 2006年第08期16卷 889-892页

作者： LIU Wei, ZHANG Ying, LIU Zhaoliang, GUO Li, WANG Xiaobo, FEI Jing, FENG Jidong, ZHAO Rui, HU Xiaoxiang and LI Ning (The State Key Laboratory for Agro-Biotechnology, College of Biology, china agricultural university, beijing 100094. china College of Animal Science and Technology, china agricultural university, beijing 100094. china) [a]The State Key Laboratory for Agno-Biotechnology College of Biology China Agricultural University Beijing China [b]College of Animal Science and Technology China Agricultural University Beijing China

A pig BAC library was constructed with genomic DNA from a male Erhualian pig. After partial digestion with HindⅢor BamHⅠthe fragments obtained were cloned into the pBeloBAC11 vector. The library consists of 184320 clones which stored in 480 pieces 384-well plates (20 plates per superpool). A two-step 4-dimension PCR screening system was established to screen the positive clones. An average insert size of 128 kb was estimated from 105 randomly isolated clones, which indicates that the library is more than five times of genomic coverage. For the demonstration of the probability to pick out any unique genes or DNA markers from the library, 10 single-copy genes were screened out and the positive clones were yielded between 1 and 8 with an average of 3.6. Positive superpools were obtained for 32 microsatellite markers selected from different regions of pig genome. The number of positive superpools for each marker varies from 1 to 9 with an average of 4.78. This BAC library provides an additional resource for pig physical mapping and gene identifica-

关键词： pig BAC library PCR screening system.

Seasonal influence on the development of parthenotes and cloned embryos in pigs

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Progress in Natural Science:Materials International 2007年第10期17卷 1172-1176页

作者： Zhang Kun1, Zhang Yunhai3,4, Pan Dengke5, Ma Yufang1, Wei Hengxi1, Zhang Jian6. Li Xuyang7, Wang Shaohua1, Li Yan2, Dai Yunping1,2 and Li Ning1(1. State Key Laboratory for Agrobiotechnology, College of Biological Science, china agricultural university, beijing 100094. china 2. JiPulin Biotech Ltd., beijing 100094. china 3. Danish Institute of agricultural Sciences, 8830 Tjele, Denmark 4. College of Animal Science and Technology, Anhui agricultural university, Hefei 230036. {6. 5. Institute of Animal Sciences, Chinese Academy of agricultural Sciences, beijing 100094. china 6. Animal Clinical Hospital, china agricultural university, beijing 100094. china 7. Hebei Minghui Group of Pigs Breeding, Sanhe 06.200, {6.) State Key Laboratory for Agrobiotechnology College of Biological Science China Agricultural University Beijing 100094 China JiPulin Biotech Ltd. Beijing 100094 China Danish Institute of agricultural Sciences 8830 Tjele Denmark College of Animal Science and Technology Anhui Agricultural University Hefei 230036 China Institute of Animal Sciences Chinese Academy of Agricultural Sciences Beijing 100094 China Animal Clinical Hospital China Agricultural University Beijing 100094 China 7. Hebei Minghui Group of Pigs Breeding Sanhe 065200 China

Seasonal influence on the developmental ability of porcine embryos was studied by comparisons of the number of cumulus-oocytes-complexes (COCs) per ovary pair and meiotic maturation ability of oocytes in different seasons; and by the observations on developmental competence of the parthenogenetic oocytes and cloned embryos collected from different seasons. We found that the number of COCs per ovary pairs was significantly higher in spring compared to the other seasons (P<0.05). However, no significant difference was found in the rate of oocytes progressing to MⅡ in four seasons (P<0.05). In addition, the rate of blastocyst formation of parthenogenetic embryos in summer was obviously declined compared to the other seasons (P<0.05), and an increased blastocyst rate of cloned porcine embryos was found in spring compared to autumn and winter (P<0.05). The results suggest that there should be a seasonal influence on the developmental competence of porcine embryos.

关键词： pig cloned parthenogenetic season